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Pluronic acid f 127

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Pluronic acid F-127 is a nonionic, water-soluble triblock copolymer. It is composed of a central hydrophobic block of polypropylene oxide (PPO) flanked by two hydrophilic blocks of polyethylene oxide (PEO). Pluronic acid F-127 exhibits thermoreversible gelation behavior, forming a clear, viscous solution at low temperatures and a semi-solid gel at higher temperatures.

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Lab products found in correlation

Fluo 4 am, by Thermo Fisher Scientific (5 mentions) Fura 2 am, by Thermo Fisher Scientific (5 mentions) Sulfinpyrazone, by Merck Group (4 mentions) Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) Fast green dye, by Merck Group (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Egta am, by Merck Group (2 mentions) Nis element software, by Nikon (2 mentions) Suramin, by Bio-Techne (1 mentions) Fluo 4, by Merck Group (1 mentions) Fluo4 am calcium dye, by Thermo Fisher Scientific (1 mentions) Xlplan n, by Olympus (1 mentions) Tryple, by Thermo Fisher Scientific (1 mentions) Poly l lysine (pll), by Merck Group (1 mentions) Fibronectin, by Merck Group (1 mentions) Cal 520 am, by AAT Bioquest (1 mentions) Carbamylcholine chloride carbachol, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Penicillin, by Merck Group (1 mentions) Potassium chloride (kcl), by Fujifilm (1 mentions)

Close spelling variants of this product

Pluronic F-127 (985 citations) Pluronic acid (29 citations) Pluronics F-127 (17 citations) Pluronic F-127 (F-127, (15 citations) Pluronic 127 (7 citations) Pluronic acid 127 (2 citations)

22 protocols using pluronic acid f 127

1

In Utero Calcium Imaging of Mouse Embryos

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Pregnant C57bl/6J mice at the 13–15th day of gestation (E13–E15) were anesthetized with intraperitoneal injection of ketamine/xylazine (80/10 mg/kg) and placed on a heating pad at 37°C.
A single 15–20 mm incision was made in the abdomen, the uterine horn exposed and embryos injected ventricularly with Fluo-4AM in loading solution using a capillary glass pipette. The loading solution consisted of 17 μg of calcium dye Fluo-4AM (Molecular Probes) dissolved in 3 μl of 20% F-127 pluronic® acid (Sigma) in dimethyl sulfoxide (DMSO; Sigma) that was then diluted in artificial cerebrospinal fluid (ACSF; in mM: 125 NaCl, 1.25 NaH2PO4, 2 CaCl2, 1 MgCl2, 5 KCl, 20 D-glucose, 10 HEPES) to reach a final concentration 0.4 mM of Fluo-4. 13 mM FastGreen dye (Sigma) was added to the solution for visual injection guidance. Injections were performed with a Picospritzer II, 1–2 μl per embryo depending on the size of the embryo through a glass pipette with a 12 mm long pulled tip that measured ~50 μm in tip diameter (Drummond Scientific) targeting the lateral ventricle. The uterine horn was rinsed with 37°C ACSF every 2 min to prevent hypothermia. After all embryos had been injected with calcium dye the uterine horn was placed back inside the mother and the cavity was sutured. In some experiments suramin (3.7 mM, Tocris Bioscience) was added to the injected solution.
Yuryev M., Pellegrino C., Jokinen V., Andriichuk L., Khirug S., Khiroug L, & Rivera C. (2016). In vivo Calcium Imaging of Evoked Calcium Waves in the Embryonic Cortex. Frontiers in Cellular Neuroscience, 9, 500.
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2

Intraventricular Calcium Imaging of Embryos

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Prior to imaging, embryos were injected intraventricularly with Fluo-4AM calcium dye (17 μg of calcium dye Fluo-4AM, Molecular Probes) dissolved in 3 μl of 20% F-127 pluronic® acid (Sigma) in dimethyl sulfoxide (DMSO, Sigma) that was then diluted in artificial cerebrospinal fluid (ACSF, in mM: 125 NaCl, 1.25 NaH2PO4, 2 CaCl2, 1 MgCl2, 5 KCl, 20 D-glucose, 10 HEPES) to reach a final concentration 0.4 mM of Fluo with 13 mM FastGreen dye (Sigma) added for injection guidance.
Two-photon imaging was performed with Olympus Fluo View 1000MP system at the rate 1.2 fps at a resolution 512 × 512 via 25X objective (XLPlan N, Olympus). Excitation light was produced by Mai-Tai DeepSee laser (Spectra Physics).
Yuryev M., Andriichuk L., Leiwe M., Jokinen V., Carabalona A, & Rivera C. (2018). In vivo two-photon imaging of the embryonic cortex reveals spontaneous ketamine-sensitive calcium activity. Scientific Reports, 8, 16059.
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3

Fluorescent Calcium Imaging with Caged IP3

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Cal520-AM was from AAT Bioquest (Sunnyvale, CA). Fluo-4-AM, Dulbecco's Modified Eagle's Medium (DMEM)/Ham's F-12 (50:50) supplemented with GlutaMAX, and TrypLE were from Invitrogen (Paisley, UK). Pluronic acid F-127, dimethyl sulfoxide (DMSO), poly-L-lysine, foetal bovine serum (FBS), histamine and carbamylcholine chloride (carbachol) were from Sigma-Aldrich (Poole, Dorset, UK). Fibronectin and EGTA-AM were from Merck Millipore (Darmstadt, Germany). Caged IP3-PM [ci-IP3-PM, d-2,3-O-isopropylidine-6-O-(2-nitro-4,5-dimethoxy)benzyl-myo-inositol 1,4,5-trisphosphate-hexakis(proprionoxymethyl)ester] was from SiChem GmbH (Bremen, Germany).
Keebler M.V, & Taylor C.W. (2017). Endogenous signalling pathways and caged IP3 evoke Ca2+ puffs at the same abundant immobile intracellular sites. Journal of Cell Science, 130(21), 3728-3739.
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4

Measurement of Intracellular Calcium Dynamics

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First passage conjunctival goblet cells were plated onto 35-mm glass bottom culture dish and incubated at 37°C overnight. Cells were then incubated for 1 hour at 37°C with KRB-HEPES plus 0.5% bovine serum albumin containing 0.5 μM fura-2/AM (Invitrogen, Grand Island, NY, USA), 8 μM pluronic acid F127 (Sigma-Aldrich, St. Louis, MO, USA) and 250 μM sulfinpyrazone (Sigma-Aldrich) for 1h. Before calcium measurement, cells were washed by KRB-HEPES containing sulfinpyrazone. Calcium measurements were conducted using a ratio imaging system (In Cyt Im2; Intracellular Imaging, Cincinnati, OH, USA) using wavelengths of 340 and 380 nm and an emission wavelength of 505 nm. Cells were stimulated with agonists, antagonists, and inhibitors. The [Ca2+]i over time was displayed and the change in peak [Ca2+]i was calculated by subtracting the average of the basal value from the peak [Ca2+]i value.
Yang M., Lippestad M., Hodges R.R., Fjærvoll H.K., Fjærvoll K.A., Bair J.A., Utheim T.P., Serhan C.N, & Dartt D.A. (2020). RvE1 Uses the LTB4 Receptor BLT1 to Increase [Ca2+]i and Stimulate Mucin Secretion in Cultured Rat and Human Conjunctival Goblet Cells. The ocular surface, 18(3), 470-482.
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5

Reagents and Concentrations for Experiments

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Chemical reagents used in the present experiment were purchased from the following sources. PPF, 5-fluoro-2’-deoxyuridine (5-FU), poly-l-lysine, streptomycin, penicillin, and Pluronic acid F-127 were obtained from Sigma−Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM), dimethyl sulfoxide (DMSO), and KCl were obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Supplement Minus Antioxidants (AO) and horse serum (HS) were obtained from Gibco BRL (Carlsbad, CA, USA) and from Dako (Carpinteria, CA, USA), respectively. Trypsin was purchased from Difco Lab (Detroit, MI, USA), fetal calf serum (FCS) from ICN Biochemicals (Costa Mesa, CA, USA), and B-27 Fluo4-AM from Dojindo (Kumamoto, Japan). PPF concentrations in the present study are comparable to the moderate concentrations prescribed for clinical use [9 (link), 17 (link)].
Shibuta S., Morita T, & Kosaka J. (2022). Effect of preconditioning on propofol-induced neurotoxicity during the developmental period. PLoS ONE, 17(8), e0273219.
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6

Fluo-4 AM Calcium Assay

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One day after seeding in 96-well plate, the cells were washed with a washing buffer (HBSS (#14065-049, Gibco), 10 mM HEPES (#15630-080, Gibco), 2 mM probenecid (#P8761, Sigma) (pH = 7.4)) and next incubated in a loading buffer (2 µM Fluo-4 AM (#F14201, Molecular Probes, in DMSO), 0.02% pluronic acid F-127 (#P2443, Sigma, in DMSO) in washing buffer) for 30 min. Cells were washed and allowed to rest for 10 min at 37°C in a washing buffer before automatic addition of BzATP. The signal was detected using the NOVOStar microplate reader system in a 2-min time span. For each data point, the maximal increase in the signal above baseline signal before BzATP addition was calculated. The resulting dose–response curves were normalized using baseline as 0% signal and signal at the highest dose as 100%. Next, the curve was fitted using a four-parameter fit of either a sigmoidal dose–response curve (one receptor) or a biphasic sigmoidal dose–response curve (two receptors). The fitted curves were finally used to calculate EC50. All calculations were performed using GraphPad Prism 7.
Dreisig K., Sund L., Dommer M.W., Kristensen N.P., Boddum K., Viste R., Fredholm S., Odum N., Jäättelä M., Skov S, & Kornum B.R. (2018). Human P2Y11 Expression Level Affects Human P2X7 Receptor-Mediated Cell Death. Frontiers in Immunology, 9, 1159.
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7

Fluorescent Calcium Imaging Assay

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Cells were stained with 5 µM Fluozin-3-AM (Invitrogen) [in SynaptoJuice B containing 0.0025% pluronic acid F127 (Sigma-Aldrich) dissolved in DMSO] for 30 min at 37˚C. Cells were washed twice and imaged in HBSS containing 1 mM CaCl2, 1 mM MgCl2 and 5 mM HEPES (pH 7.2). Images were taken on the Colibri LED microscope system at 470 nm (excitation)/510 nm (emission) or Perkin Elmer Operetta high-content imaging system.
Somogyi A., Kirkham E.D., Lloyd-Evans E., Winston J., Allen N.D., Mackrill J.J., Anderson K.E., Hawkins P.T., Gardiner S.E., Waller-Evans H., Sims R., Boland B, & O'Neill C. (2023). The synthetic TRPML1 agonist ML-SA1 rescues Alzheimer-related alterations of the endosomal-autophagic-lysosomal system. Journal of Cell Science, 136(6), jcs259875.
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8

Calcium Imaging of ESCs Stimulated by Adenosine

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Undifferentiated and differentiated E14Tg2A ESCs were loaded with 5 µM of Fluo3-AM for 45 min at 37°C in DMEM high glucose in 0.5% Me2SO and 0.06% of the nonionic surfactant pluronic acid F-127 (Sigma-Aldrich). After loading with Fluo-3 AM, the cells were incubated with extracellular buffer (140 mM NaCl, 3 mM KCl, 1 mM MgCl2, 2 mM CaCl2, 10 mM HEPES, and 10 mM glucose at pH 7.4) (Glaser et al., 2020 (link)). Ca2+ imaging was performed using the Inverted Research Microscope ECLIPSE-TiS (Nikon, Melville, NY) equipped with a 14-bit high-resolution CoolSNAP HQ2 CCD camera (Photometrics, Tucson, AZ) and analyzed with the NISElement software (Nikon) using image acquisition rates of two frames per second. Fluo-3 fluorescence was excited with a xenon lamp at 488 nm, and the emitted light was detected using a bandpass filter at 515–530 nm. Intracellular calcium flux was monitored in cells stimulated with adenosine (2 µM).
Glaser T., Martins P., Beco R., Bento C.A., Cappellari A.R., La Banca Oliveira S., Merkel C.A., Arnaud-Sampaio V.F., Lameu C., Battastini A.M, & Ulrich H. (2024). Impairment of adenosine signaling disrupts early embryo development: unveiling the underlying mechanisms. Frontiers in Pharmacology, 14, 1328398.
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9

Intracellular Calcium Imaging in ESCs

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Undifferentiated and differentiated E14Tg2A ESC were loaded with 5 µM of Fluo3-AM for 45 min at 37°C in DMEM High glucose in 0.5% Me2SO and 0.06% of the nonionic surfactant pluronic acid F-127 (Sigma Aldrich). After loading with Fluo-3AM, the cells were incubated with extracellular buffer (140 mM NaCl, 3 mM KCl, 1 mM MgCl2, 2 mM CaCl2, 10 mM HEPES, 10 mM glucose at pH 7.4) [25] (link). Ca2+ imaging was performed by using the Inverted Research Microscope ECLIPSE-TiS (Nikon, Melville, NY) equipped with a 14 bit high-resolution CCD camera CoolSNAP HQ2 (Photometrics, Tucson, AZ) and analyzed with NIS-Element software (Nikon) using image acquisition rates of two frames per second. Fluo-3 fluorescence was excited with a xenon lamp at 488 nm, and the emitted light was detected using a band-pass filter at 515–530 nm. Intracellular calcium influx was monitored in cells stimulated with 10 µM ATP and 10 µM Bz-ATP. Forty cells were analyzed for each data point and the calcium influx was determined as mean variation between Fluo-3AM fluorescence intensities obtained during the stimulus (F) and the rest state (Fo), normalized by its basal fluorescence (Fb). (F-Fo)/Fb.
Glaser T., de Oliveira S.L., Cheffer A., Beco R., Martins P., Fornazari M., Lameu C., Junior H.M., Coutinho-Silva R, & Ulrich H. (2014). Modulation of Mouse Embryonic Stem Cell Proliferation and Neural Differentiation by the P2X7 Receptor. PLoS ONE, 9(5), e96281.
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10

Ulvan-based Hydrogel Biomaterial Synthesis

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Ulvan polysaccharide powder extracted by Ulva armoricana was provided by ELICITYL (France) (molecular weight, from 90 to 500 kDa). Microcrystalline cellulose powder (MCC), sodium periodate (NaIO4), hyaluronic acid (Mw = 1.5-1.8 MDa), Pluronic acid F-127, Bovine Serum Albumin, 2-Hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone (Irgacure D-2959), methacrylic anhydride and phosphate buffered saline (PBS) were all purchased from Sigma-Aldrich, Missouri, St. Louis, (USA). Sodium hydroxide and hydrochloric acid were purchased from VWR, Fontenay-sous-Bois (France). Concentrated sulfuric acid (95−98%) was purchased from Laborspirit, Santo Antão do Tojal (Portugal). Sodium hyaluronate (Mw = 253 kDa) purchased from Lifecore Biomedical, Minnesota, Chaska (USA).
Massironi A., Franco A.R., Babo P.S., Puppi D., Chiellini F., Reis R.L, & Gomes M.E. (2022). Development and Characterization of Highly Stable Silver NanoParticles as Novel Potential Antimicrobial Agents for Wound Healing Hydrogels. International Journal of Molecular Sciences, 23(4), 2161.
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